lncRNA RPL22P1-201通过调控miR-216b-5p表达影响前列腺癌细胞增殖、细胞周期和多西紫杉醇的敏感性

LncRNA RPL22P1-201 affects prostate cancer cell proliferation,cell cycle,and sensitivity to docetaxel by regulating miR-216b-5p expression

目的:探究长链非编码RNA(lncRNA)RPL22P1-201通过介导miR-216b-5p表达对前列腺癌细胞增殖、细胞周期以及多西紫杉醇敏感性的作用机制。方法:基于Cancer LncRNA Census数据库分析前列腺癌组织和正常组织中RPL22P1-201的表达差异。实时定量聚合酶链式反应(qRT-PCR)检测前列腺癌细胞株(DU-145、C4-2B、PC3、22Rv1、LNCaP)和正常前列腺上皮细胞(RWPE-1)中RPL22P1-201表达水平。将PC3细胞分为si-RPL22P1-201组(转染RPL22P1-201干扰序列)和si-NC组(转染si-NC序列),集落形成实验检测PC3细胞增殖能力,流式细胞术检测PC3细胞周期,CCK-8法检测多西紫杉醇处理后各组PC3细胞的增殖情况,双荧光素酶报告基因实验验证RPL22P1-201与靶基因的结合,qRT-PCR检测miR-216b-5p表达水平,Western印迹检测TrkB、CDK4、cyclin D2、cyclin D3、CDK6蛋白表达水平。结果:前列腺癌组织中RPL22P1-201表达水平高于正常组织(P<0.01),前列腺癌细胞株中RPL22P1-201表达水平高于正常前列腺上皮细胞(P<0.01),si-NC组和si-RPL22P1-201组集落数量分别为(256.1±28.79)个和(78.77±14.52)个,差异有统计学意义(P<0.01)。si-NC组和si-RPL22P1-201组G0/G1细胞率分别为(43.18±4.56)%和(68.85±3.40)%,S细胞率分别为(36.84±2.28)%和(24.27±2.74)%,G2/M细胞率分别为(19.98±2.69)%和(6.88±1.57)%,差异均有统计学意义(均P<0.05)。si-RPL22P1-201组在多西紫杉醇作用下的细胞存活率均低于si-NC组(均P<0.05),RPL22P1-201能够与miR-216b-5p配对结合(P<0.01)。相比si-NC组,si-RPL22P1-201组PC3细胞中miR-216b-5p表达降低(P<0.01),TrkB、CDK4、cyclin D2、cyclin D3、CDK6蛋白表达均降低。结论:RPL22P1-201在前列腺癌中高表达,沉默RPL22P1-201通过升高miR-216b-5p表达抑制前列腺癌PC3细胞增殖和细胞周期,并增强PC3细胞对多西紫杉醇的敏感性。

Objective:Exploring the effects and mechanisms of long non coding RNA(IncRNA)RPL22P1-201 on prostate cancer cell proliferation,cell cycle,and docetaxel sensitivity by regulating miR-216b-5p expression.Methods:The Cancer LncRNA Census database was used to analyze the differential expression of RPL22P1-201 between prostate cancer tissue and normal tissue.Real time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression level of RPL22P1-201 in prostate cancer cell lines(DU-145,C4-2B,PC3,22Rvl,LNCaP)and normal prostate epithelial cells(RWPE-1).PC3 cells were divided into si-RPL22P1-201 group(transfected with RPL22P1-201 interference sequence)and si-NC group(transfected with si-NC sequence).Colony formation assay was used to detect the proliferation ability of PC3 cells.Flow cytometry was used to detect the PC3 cell cycle.The CCK-8 method was used to detect the proliferation of PC3 cells in each group after treatment with docetaxel.The dual luciferase reporter gene experiment verifies the binding of RPL22P1-201 to the target gene.qRT-PCR was used to detect the expression level of miR-216b-5p.Western blot was used to detect the expression levels of TrkB,CDK4,cyclin D2,cyclin D3,and CDK6 proteins.Results:The expression level of RPL22P1-201 in prostate cancer tissue was higher than that in normal tissue(P<0.01).The expression level of RPL22P1-201 in prostate cancer cell lines was higher than that in normal prostate epithelial cells(P<0.01).The number of colonies in the si-NC group and si-RPL22Pl-201 group was(256.1±28.79)and(78.77±14.52),respectively.The difference was statistically significant(P<0.01).The G0/G1 cell rates in the si-NC group and si-RPL22Pl-201 group were(43.18±4.56)%and(68.85±3.40)%,respectively.The S cell rates were(36.84±2.28)%and(24.27±2.74)%,respectively.The G2/M cell rates were(19.98±2.69)%and(6.88±1.57)%,respectively,and the differences were statistically significant(all P<0.05).The cell survival rate of the si-RPL22Pl-201 group under the action of docetaxel was lower than that of the si-NC group(all P<0.05).RPL22P1-201 can pair and bind with miR-216b-5p(P<0.01).Compared with the si-NC group,the si-RPL22P1-201 group showed a decrease in miR-216b-5p expression in PC3 cells(P<0.01),and a decrease in TrkB,CDK4,cyclin D2,cyclin D3,and CDK6 protein expression.Conclusions:RPL22P1-201 is highly expressed in prostate cancer,and silencing RPL22P1-201 inhibits prostate cancer PC3 cell proliferation and cell cycle by increasing miR-216b-5p expression,and enhances PC3 cell sensitivity to docetaxel.